Difference between revisions of "Sandbox"

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−
* '''Description:''' aminomethyltransferase (glycine cleavage system protein T) <br/><br/>
+
* '''Description:''' Transcriptional antiterminator, controls expression of the ''ptsGHI'' operon<br/><br/>
  
 
{| align="right" border="1" cellpadding="2"  
 
{| align="right" border="1" cellpadding="2"  
 
|-
 
|-
 
|style="background:#ABCDEF;" align="center"|'''Gene name'''
 
|style="background:#ABCDEF;" align="center"|'''Gene name'''
−
|''gcvT''
+
|''glcT''
 
|-
 
|-
−
|style="background:#ABCDEF;" align="center"| '''Synonyms''' || ''yqhI ''
+
|style="background:#ABCDEF;" align="center"| '''Synonyms''' || ''ykwA''
 
|-
 
|-
−
|style="background:#ABCDEF;" align="center"| '''Essential''' || no  
+
|style="background:#ABCDEF;" align="center"| '''Essential''' || no
 
|-
 
|-
−
|style="background:#ABCDEF;" align="center"| '''Product''' || aminomethyltransferase <br/>(glycine cleavage system protein T)
+
|style="background:#ABCDEF;" align="center"| '''Product''' || transcriptional antiterminator of the ''ptsGHI'' operon
 
|-
 
|-
−
|style="background:#ABCDEF;" align="center"|'''Function''' || glycine utilization
+
|style="background:#ABCDEF;" align="center"|'''Function''' || transcriptional antiterminator
 
|-
 
|-
−
|style="background:#ABCDEF;" align="center"| '''MW, pI''' || 39 kDa, 5.734 
+
|style="background:#ABCDEF;" align="center"| '''MW, pI''' || 33,0 kDa, 7.01
 
|-
 
|-
−
|style="background:#ABCDEF;" align="center"| '''Gene length, protein length''' || 1086 bp, 362 aa
+
|style="background:#ABCDEF;" align="center"| '''Gene length, protein length''' || 855 bp, 285 amino acids
 
|-
 
|-
−
|style="background:#ABCDEF;" align="center"|'''Immediate neighbours''' || ''[[gcvPA]]'', ''[[yqhH]]''
+
|style="background:#ABCDEF;" align="center"|'''Immediate neighbours''' || ''[[ykvZ]]'', ''[[ptsG]]''
 
|-
 
|-
−
|colspan="2" style="background:#FAF8CC;" align="center"|'''Get the DNA and protein [http://srs.ebi.ac.uk/srsbin/cgi-bin/wgetz?-e+&#91;EMBLCDS:CAB14388&#93;+-newId sequences] <br/> (Barbe ''et al.'', 2009)'''
+
|colspan="2" style="background:#FAF8CC;" align="center"|'''Get the DNA and protein [http://srs.ebi.ac.uk/srsbin/cgi-bin/wgetz?-e+&#91;EMBLCDS:CAB13261&#93;+-newId sequences] <br/> (Barbe ''et al.'', 2009)'''
 
|-
 
|-
−
|-
+
|colspan="2" | '''Genetic context''' <br/> [[Image:glcT_context.gif]]
−
|-
 
−
|colspan="2" | '''Genetic context''' <br/> [[Image:gcvT_context.gif]]
 
 
  <div align="right"> <small>This image was kindly provided by [http://genolist.pasteur.fr/SubtiList/ SubtiList]</small></div>
 
  <div align="right"> <small>This image was kindly provided by [http://genolist.pasteur.fr/SubtiList/ SubtiList]</small></div>
 
|-
 
|-
Line 31: Line 29:
 
__TOC__
 
__TOC__
  
−
<br/><br/><br/><br/><br/>
+
<br/><br/>
 +
 
  
 
=The gene=
 
=The gene=
Line 37: Line 36:
 
=== Basic information ===
 
=== Basic information ===
  
−
* '''Locus tag:''' BSU24570
+
* '''Locus tag:''' BSU13880
  
 
===Phenotypes of a mutant ===
 
===Phenotypes of a mutant ===
Line 43: Line 42:
 
=== Database entries ===
 
=== Database entries ===
  
−
* '''DBTBS entry:''' [http://dbtbs.hgc.jp/COG/prom/gcvT-gcvPA-gcvPB.html]
+
* '''DBTBS entry:''' no entry
  
−
* '''SubtiList entry:''' [http://genolist.pasteur.fr/SubtiList/genome.cgi?gene_detail+BG11509]
+
* '''SubtiList entry:''' [http://genolist.pasteur.fr/SubtiList/genome.cgi?gene_detail+BG12593]
  
 
=== Additional information===
 
=== Additional information===
−
 
  
 
=The protein=
 
=The protein=
Line 54: Line 52:
 
=== Basic information/ Evolution ===
 
=== Basic information/ Evolution ===
  
−
* '''Catalyzed reaction/ biological activity:''' [Protein]-S(8)-aminomethyldihydrolipoyllysine + tetrahydrofolate = [protein]-dihydrolipoyllysine + 5,10-methylenetetrahydrofolate + NH<sub>3</sub> (according to Swiss-Prot)
+
* '''Catalyzed reaction/ biological activity:''' transcriptional antiterminator, RNA-binding protein, binds the ''[[ptsG]]'' RAT sequence
  
−
* '''Protein family:''' gcvT family (according to Swiss-Prot)
+
* '''Protein family:''' antiterminator of the BglG/ [[SacY]] family
  
−
* '''Paralogous protein(s):'''
+
* '''Paralogous protein(s):''' [[LicT]], [[SacT]], [[SacY]]
  
 
=== Extended information on the protein ===
 
=== Extended information on the protein ===
Line 65: Line 63:
  
 
* '''Domains:'''  
 
* '''Domains:'''  
 +
** RNA-binding domain (N-terminal, constitutive antiterminator)
 +
** 2x PTS regulation domains (PRDs) (C-terminal, neg. regulated by [[PtsG]])
  
−
* '''Modification:'''
+
* '''Modification:''' phosphorylation (His104)
  
 
* '''Cofactor(s):'''
 
* '''Cofactor(s):'''
Line 72: Line 72:
 
* '''Effectors of protein activity:'''
 
* '''Effectors of protein activity:'''
  
−
* '''Interactions:'''
+
* '''Interactions:''' [[PtsH]]-[[GlcT]]
  
 
* '''Localization:'''
 
* '''Localization:'''
Line 78: Line 78:
 
=== Database entries ===
 
=== Database entries ===
  
−
* '''Structure:''' [http://www.rcsb.org/pdb/explore.do?structureId=1YX2 1YX2]
+
* '''Structure:'''
  
−
* '''Swiss prot entry:''' [http://www.uniprot.org/uniprot/P54378 P54378]
+
* '''Swiss prot entry:''' [http://www.uniprot.org/uniprot/O31691]
  
−
* '''KEGG entry:''' [http://www.genome.jp/dbget-bin/www_bget?bsu+BSU24570]
+
* '''KEGG entry:''' [http://www.genome.jp/dbget-bin/www_bget?bsu+BSU13880]
  
−
* '''E.C. number:''' [http://www.expasy.org/enzyme/2.1.2.10 2.1.2.10]
+
=== Additional information===
  
−
=== Additional information===
 
  
 
=Expression and regulation=
 
=Expression and regulation=
  
−
* '''Operon:''''' [[gcvT]]-[[gcvPA]]-[[gcvPB]]'' [http://www.ncbi.nlm.nih.gov/sites/entrez/15472076 PubMed]
+
* '''Operon:'''  
  
−
* '''[[Sigma factor]]:'''  
+
* '''Sigma factor:'''  
  
−
* '''Regulation:''' induced by glycine [http://www.ncbi.nlm.nih.gov/sites/entrez/15472076 PubMed]
+
* '''Regulation:'''  
  
−
* '''Regulatory mechanism:''' induction requires a co-operatively acting riboswitch [http://www.ncbi.nlm.nih.gov/sites/entrez/15472076 PubMed]
+
* '''Regulatory mechanism:'''  
  
−
* '''Additional information:'''
+
* '''Additional information:'''  
  
 
=Biological materials =
 
=Biological materials =
  
−
* '''Mutant:'''
+
* '''Mutant:''' GP109 (in frame deletion), available in [[Stülke]] lab
  
−
* '''Expression vector:'''
+
* '''Expression vector:''' pGP124 (full length, in [[pWH844]]), pGP114 (amino acids 1-60, RNA-binding domain, in [[pWH844]]), pGP230 (amino acids 1-60, RNA-binding domain with thrombin cleavage site, in [[pWH844]]), pGP164 (both PRDs, in [[pWH844]]), in addition diverse expression vectors for phosphorylation site mutants and for RBD mutants (all in [[pWH844]]), pGP424 (PRDI, in [[pWH844]]), pGP425 (PRDII, in [[pWH844]]), pGP442 (PRDI, in [[pGP570]], with thrombin cleavage site), pGP443 (PRDII, in [[pGP570]],  with thrombin cleavage site), pGP575 (amino acids 1-60, RNA-binding domain with Strep-tag, in [[pGP574]]), all available in [[Stülke]] lab
−
       
+
 
* '''lacZ fusion:'''
 
* '''lacZ fusion:'''
  
 
* '''GFP fusion:'''
 
* '''GFP fusion:'''
−
 
−
* '''two-hybrid system:'''
 
  
 
* '''Antibody:'''
 
* '''Antibody:'''
  
 
=Labs working on this gene/protein=
 
=Labs working on this gene/protein=
 +
 +
[[Stülke|Jörg Stülke]], University of Göttingen, Germany
 +
[http://wwwuser.gwdg.de/~genmibio/stuelke.html Homepage]
  
 
=Your additional remarks=
 
=Your additional remarks=
Line 120: Line 120:
 
=References=
 
=References=
  
−
<pubmed>15472076, </pubmed>
+
<pubmed>11902727 9765562 12437213 10543968 17074746 15155854 14527945 9663674, </pubmed>
−
# Mandal M, Lee M, Barrick JE, Weinberg Z, Emilsson GM, Ruzzo WL, Breaker RR (2004) A glycine-dependent riboswitch that uses cooperative binding to control gene expression. ''Science'' '''306:''' 275-279. [http://www.ncbi.nlm.nih.gov/sites/entrez/15472076 PubMed]
+
# Stülke J, Martin-Verstraete I, Zagorec M (1997) Induction of the ''Bacillus subtilis ptsGHI'' operon by glucose is controlled by a novel antiterminator, ''GlcT Mol Microbiol.'' '''25:''' 65-78. [http://www.ncbi.nlm.nih.gov/sites/entrez/11902727 PubMed]
−
# Author1, Author2 & Author3 (year) Title ''Journal'' '''volume:''' page-page. [http://www.ncbi.nlm.nih.gov/sites/entrez/PMID PubMed]
+
# Bachem S, Stülke J. (1998)   Regulation of the ''Bacillus subtilis'' GlcT antiterminator protein by components of the phosphotransferase system. ''J Bacteriol.'' '''180:''' 5319-26 [http://www.ncbi.nlm.nih.gov/sites/entrez/9765562 PubMed]
 +
# Greenberg, D. B., Stülke, J. & Saier Jr., M. H. (2002) Domain analysis of transcriptional regulators bearing PTS-regulatory domains. Res. Microbiol. 153: 519-526. [http://www.ncbi.nlm.nih.gov/sites/entrez/12437213 PubMed]
 +
# Langbein, I., Bachem, S. & Stülke, J. (1999) Specific interaction of the RNA binding domain of the Bacillus subtilis transcriptional antiterminator GlcT with its RNA target, RAT. J. Mol. Biol. 293: 795-805. [http://www.ncbi.nlm.nih.gov/sites/entrez/10543968 PubMed]
 +
# Schilling, O., Herzberg, C., Hertrich, T., Vörsmann, H., Jessen, D., Hübner, S., Titgemeyer, F. & Stülke, J. (2006) Keeping signals straight in transcription regulation: specificity determinants for the interaction of a family of conserved bacterial RNA-protein couples. Nucl. Acids Res. 34: 6102-6115. [http://www.ncbi.nlm.nih.gov/sites/entrez/17074746 PubMed]
 +
# Schilling, O., Langbein, I., Müller, M., Schmalisch, M. & Stülke, J. (2004) A protein-dependent riboswitch controlling ptsGHI operon expression in Bacillus subtilis: RNA structure rather than sequence provides interaction specificity. Nucl. Acids Res. 32: 2853-2864. [http://www.ncbi.nlm.nih.gov/sites/entrez/15155854 PubMed]
 +
# Schmalisch, M., Bachem, S. & Stülke, J. (2003) Control of the Bacillus subtilis antiterminator protein GlcT by phosphorylation: Elucidation of the phosphorylation chain leading to inactivation of GlcT. J. Biol. Chem. 278: 51108-51115. [http://www.ncbi.nlm.nih.gov/sites/entrez/14527945 PubMed]
 +
# Stülke, J., Arnaud, M., Rapoport, G., & Martin-Verstraete, I. (1998) PRD - A protein domain involved in PTS-dependent induction and carbon catabolite repression of catabolic operons in bacteria. Mol. Microbiol. 28: 865-874. [http://www.ncbi.nlm.nih.gov/sites/entrez/9663674 PubMed]

Revision as of 14:04, 8 June 2009

  • Description: Transcriptional antiterminator, controls expression of the ptsGHI operon

Gene name glcT
Synonyms ykwA
Essential no
Product transcriptional antiterminator of the ptsGHI operon
Function transcriptional antiterminator
MW, pI 33,0 kDa, 7.01
Gene length, protein length 855 bp, 285 amino acids
Immediate neighbours ykvZ, ptsG
Get the DNA and protein sequences
(Barbe et al., 2009)
Genetic context
GlcT context.gif
This image was kindly provided by SubtiList




The gene

Basic information

  • Locus tag: BSU13880

Phenotypes of a mutant

Database entries

  • DBTBS entry: no entry
  • SubtiList entry: [1]

Additional information

The protein

Basic information/ Evolution

  • Catalyzed reaction/ biological activity: transcriptional antiterminator, RNA-binding protein, binds the ptsG RAT sequence
  • Protein family: antiterminator of the BglG/ SacY family

Extended information on the protein

  • Kinetic information:
  • Domains:
    • RNA-binding domain (N-terminal, constitutive antiterminator)
    • 2x PTS regulation domains (PRDs) (C-terminal, neg. regulated by PtsG)
  • Modification: phosphorylation (His104)
  • Cofactor(s):
  • Effectors of protein activity:
  • Localization:

Database entries

  • Structure:
  • Swiss prot entry: [2]
  • KEGG entry: [3]

Additional information

Expression and regulation

  • Operon:
  • Sigma factor:
  • Regulation:
  • Regulatory mechanism:
  • Additional information:

Biological materials

  • Mutant: GP109 (in frame deletion), available in Stülke lab
  • Expression vector: pGP124 (full length, in pWH844), pGP114 (amino acids 1-60, RNA-binding domain, in pWH844), pGP230 (amino acids 1-60, RNA-binding domain with thrombin cleavage site, in pWH844), pGP164 (both PRDs, in pWH844), in addition diverse expression vectors for phosphorylation site mutants and for RBD mutants (all in pWH844), pGP424 (PRDI, in pWH844), pGP425 (PRDII, in pWH844), pGP442 (PRDI, in pGP570, with thrombin cleavage site), pGP443 (PRDII, in pGP570, with thrombin cleavage site), pGP575 (amino acids 1-60, RNA-binding domain with Strep-tag, in pGP574), all available in Stülke lab
  • lacZ fusion:
  • GFP fusion:
  • Antibody:

Labs working on this gene/protein

Jörg Stülke, University of Göttingen, Germany Homepage

Your additional remarks

References

Oliver Schilling, Christina Herzberg, Tina Hertrich, Hanna Vörsmann, Dirk Jessen, Sebastian Hübner, Fritz Titgemeyer, Jörg Stülke
Keeping signals straight in transcription regulation: specificity determinants for the interaction of a family of conserved bacterial RNA-protein couples.
Nucleic Acids Res: 2006, 34(21);6102-15
[PubMed:17074746] [WorldCat.org] [DOI] (I p)

Oliver Schilling, Ines Langbein, Michael Müller, Matthias H Schmalisch, Jörg Stülke
A protein-dependent riboswitch controlling ptsGHI operon expression in Bacillus subtilis: RNA structure rather than sequence provides interaction specificity.
Nucleic Acids Res: 2004, 32(9);2853-64
[PubMed:15155854] [WorldCat.org] [DOI] (I e)

Matthias H Schmalisch, Steffi Bachem, Jörg Stülke
Control of the Bacillus subtilis antiterminator protein GlcT by phosphorylation. Elucidation of the phosphorylation chain leading to inactivation of GlcT.
J Biol Chem: 2003, 278(51);51108-15
[PubMed:14527945] [WorldCat.org] [DOI] (P p)

David B Greenberg, Jorg Stülke, Milton H Saier
Domain analysis of transcriptional regulators bearing PTS regulatory domains.
Res Microbiol: 2002, 153(8);519-26
[PubMed:12437213] [WorldCat.org] [DOI] (P p)

I Langbein, S Bachem, J Stülke
Specific interaction of the RNA-binding domain of the bacillus subtilis transcriptional antiterminator GlcT with its RNA target, RAT.
J Mol Biol: 1999, 293(4);795-805
[PubMed:10543968] [WorldCat.org] [DOI] (P p)

S Bachem, J Stülke
Regulation of the Bacillus subtilis GlcT antiterminator protein by components of the phosphotransferase system.
J Bacteriol: 1998, 180(20);5319-26
[PubMed:9765562] [WorldCat.org] [DOI] (P p)

J Stülke, M Arnaud, G Rapoport, I Martin-Verstraete
PRD--a protein domain involved in PTS-dependent induction and carbon catabolite repression of catabolic operons in bacteria.
Mol Microbiol: 1998, 28(5);865-74
[PubMed:9663674] [WorldCat.org] [DOI] (P p)

J Stülke, I Martin-Verstraete, M Zagorec, M Rose, A Klier, G Rapoport
Induction of the Bacillus subtilis ptsGHI operon by glucose is controlled by a novel antiterminator, GlcT.
Mol Microbiol: 1997, 25(1);65-78
[PubMed:11902727] [WorldCat.org] [DOI] (P p)

  1. Stülke J, Martin-Verstraete I, Zagorec M (1997) Induction of the Bacillus subtilis ptsGHI operon by glucose is controlled by a novel antiterminator, GlcT Mol Microbiol. 25: 65-78. PubMed
  2. Bachem S, Stülke J. (1998) Regulation of the Bacillus subtilis GlcT antiterminator protein by components of the phosphotransferase system. J Bacteriol. 180: 5319-26 PubMed
  3. Greenberg, D. B., Stülke, J. & Saier Jr., M. H. (2002) Domain analysis of transcriptional regulators bearing PTS-regulatory domains. Res. Microbiol. 153: 519-526. PubMed
  4. Langbein, I., Bachem, S. & Stülke, J. (1999) Specific interaction of the RNA binding domain of the Bacillus subtilis transcriptional antiterminator GlcT with its RNA target, RAT. J. Mol. Biol. 293: 795-805. PubMed
  5. Schilling, O., Herzberg, C., Hertrich, T., Vörsmann, H., Jessen, D., Hübner, S., Titgemeyer, F. & Stülke, J. (2006) Keeping signals straight in transcription regulation: specificity determinants for the interaction of a family of conserved bacterial RNA-protein couples. Nucl. Acids Res. 34: 6102-6115. PubMed
  6. Schilling, O., Langbein, I., Müller, M., Schmalisch, M. & Stülke, J. (2004) A protein-dependent riboswitch controlling ptsGHI operon expression in Bacillus subtilis: RNA structure rather than sequence provides interaction specificity. Nucl. Acids Res. 32: 2853-2864. PubMed
  7. Schmalisch, M., Bachem, S. & Stülke, J. (2003) Control of the Bacillus subtilis antiterminator protein GlcT by phosphorylation: Elucidation of the phosphorylation chain leading to inactivation of GlcT. J. Biol. Chem. 278: 51108-51115. PubMed
  8. Stülke, J., Arnaud, M., Rapoport, G., & Martin-Verstraete, I. (1998) PRD - A protein domain involved in PTS-dependent induction and carbon catabolite repression of catabolic operons in bacteria. Mol. Microbiol. 28: 865-874. PubMed
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